Growth hormone (GH) and GH-releasing hormone (GHRH): Co-localization and action in the chicken testis

• GH and GHRH co-localize in chicken testes, mainly in spermatogonia and spermatocytes.
• GHRH induced a dose-dependent increase in GH release in testicular cell cultures.
• 15 kDa GH isoforms were more abundant in culture media after GHRH stimulation.
• GHRH treatment also increased testicular PCNA immunoreactivity in the testes.
• GH increased proliferation and metabolic activity of testicular cells.

Growth hormone (GH) gene expression is not confined to the pituitary gland and occurs in many extrapituitary tissues, including the chicken testis. The regulation and function of GH in extrapituitary tissues is, however, largely unknown. The possibility that chicken testicular GH might be regulated by GH-releasing hormone (GHRH), as in the avian pituitary gland, was investigated in the present study. GHRH co-localized with GH in the germinal epithelium and in interstitial zones within the chicken testes, particularly in the spermatogonia and spermatocytes. In testicular cell cultures, exogenous human GHRH1–44 induced (at 1, 10 and 100 nM) a dose-related increase in GH release. Western blot analysis showed a heterogeneous pattern in the GH moieties released during GHRH stimulation. 26 kDa monomer GH was the most abundant moiety under basal conditions, but 15 and 17 kDa isoforms were more abundant after GHRH stimulation. GHRH treatment also increased the abundance of PCNA (proliferating cell nuclear antigen) immunoreactivity in the testes. This may have been GH-mediated, since exogenous GH similarly increased the incorporation of (3H)-thymidine into cultured testicular cells and increased their metabolic activity, as determined by increased MTT reduction. Furthermore, GH and GHRH immunoneutralization blocked GHRH-stimulated proliferative activity. In summary, these results indicate that GHRH stimulates testicular GH secretion in an autocrine or paracrine manner. Data also demonstrate proliferative actions of GHRH on testicular cell number and suggest that this action is mediated by local GH production.